Protein Isolation and Analysis » His Tag ELISA Detection Kit
His Tag ELISA Detection Kit

His Tag ELISA Detection Kit

Concentration gradient represents X, OD450 value represents Y, all X values will be transformed to Log(X) to create the standard curve which was calculated by 4PL (Figure 1). The operator can get log values of sample concentration by inputting the OD450 value of unknown samples. By formula X=10X, sample concentration can be calculated.

His Tag ELISA Detection Kit

GenScript His Tag ELISA Detection Kit, a 1.5 hour competition ELISA, is developed for rapid and high throughput detection of His-tagged proteins.
L00436
¥41,720.00

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Note

L00436 interferes with IgG in mouse/rat serum and is not suitable for detecting mouse/rat serum samples.

Overview
Product Description His tag is successive histidine (H) residues and there are mainly three forms: HHHHHH (6 x His), HHHHH (5 x His), HHHH (4 x His). Due to its small size, less interfere in protein folding, weak immunogenicity, His tag is the most dominant tag, which is widely used in recombinant protein expression. The DNA sequence which codes for His tag, is usually constructed at N-terminus or C-terminus of variety of expression plasmids. Since His tag has high affinity for Ni2+ ions, His-tagged proteins can be easily purified from bacteria, yeast and mammalian cell expression system by Ni2+-resin chromatography. Anti-His tag antibody is a useful tool for the identification of His-tagged proteins with various methods such as western blot, immunoprecipitation and flow cytometry.
GenScript His Tag ELISA Detection Kit, a 1.5 hour competition ELISA, is developed for rapid and high throughput detection of His-tagged proteins. There are several potential usages:
●Quickly identify the presence of His-tagged proteins in samples.
●Optimize protein expression by monitoring the His-tagged proteins level.
●High throughput screening of stable cell lines expressing His-tagged proteins.
This kit is based on competitive ELISA method. His Tag Plate in the kit is a 96-well microtiter plate coated with a His-tagged protein with the molecular weight of 12.7 kDa. The plate, which comprises of 8 wells x 12 strips, is demountable. When Anti-His Monoclonal Antibody and His-tagged proteins are added to the well , the coated His-tagged proteins compete with His Tag Standard (His-tagged protein with the molecular weight of 11.3 kDa) in solution or His-tagged protein in sample to interact with Anti-His Monoclonal Antibody. The higher the concentration of the His-tagged protein in solution is, the less the antibody bound to the plate will be. The Anti-His Monoclonal Antibody used in the kit is a mouse anti His tag monoclonal antibody (GenScript A00186). Antibody Tracer, which is a horseradish peroxidase (HRP) conjugated Goat anti mouse IgG, is used for enzyme reaction. His-tagged protein, Anti-His Monoclonal Antibody and antibody tracer form a complex. Other unbound molecules can be removed by washing solution. The antibody tracer reacts with TMB substrate to develop blue product that turns yellow immediately when the Stop Solution is added, which can be measured by microplate reader at 450 nM.
In optimized test condition, each absorbance value is indicated to the individual His-tagged proteins amount in solution.  His-tagged protein standards of known concentration and the corresponding absorbance values are used to construct a standard curve. With the standard curve, His-tagged protein amount present in the unknown sample can be calculated by transforming its absorbance value. 

Product Introduction
Sensitivity 1 ng/ml His-tagged proteins
Detection Range 1 ng/ml~729 ng/ml
Test Samples N-terminal/C-terminal/internal His-tagged proteins4 x His/5 x His/6 x His-tagged proteins
Mammalian, yeast and bacteria cell lysates or cell supernatant
Conveniency Provide all reagents required for test
Complete test within 1.5 hours
Kit Contents
Component Quantity Part. No
His Tag Plate 1 plate (8 wells x 12 strips) 436-80
Anti-His Monoclonal Antibody 6 ml 436-20
Antibody Tracer 12 ml 436-30
His Tag Standards
(0,1,3,9,27,81,243,729 ng/ml)		 1 ml per each standard 436-11,12,13,14,15,16,17,18
Assay Diluent 60 ml 436-60
20 × Wash Solution 40 ml 436-70
TMB Substrate 12 ml 436-40
Stop Solution 6 ml 436-50
His Tag Standard Stock (10 μg/ml) 500 μl 436-10
Plate Sealer 2 N/A
User Manual 1 N/A
Reagents compatibility Tolerable with various reagents at certain concentration
Storage The unopened kit is stable for at least 12 months if stored at 2-8 °C and the opened kit may be stable for up to 1 month at 2-8 °C. Do not freeze the kit.

Examples
  • His Tag ELISA Detection Kit
    • His Tag ELISA Detection Kit

  • His Tag ELISA Detection Kit
    • His Tag ELISA Detection Kit

    Concentration gradient represents X, OD450 value represents Y, all X values will be transformed to Log(X) to create the standard curve which was calculated by 4PL (Figure 1). The operator can get log values of sample concentration by inputting the OD450 value of unknown samples. By formula X=10X, sample concentration can be calculated.


For Research Use Only. Not for Use in Diagnostic Procedures.